A retrovirus designated human immunodeficiency virus (HIV) is the etiological agent of the complex disease that includes progressive destruction of the immune system (acquired immune deficiency syndrome; AIDS) and degeneration of the central and peripheral nervous system. This virus was previously known as LAV, HTLV-III, or ARV. A common feature of retrovirus replication is the insertion by virally-encoded integrase of proviral DNA into the host cell genome, a required step in HIV replication in human T-lymphoid and monocytoid cells. Integration is believed to be mediated by integrase in three steps: assembly of a stable nucleoprotein complex with viral DNA sequences; cleavage of two nucleotides from the 3' termini of the linear proviral DNA; covalent joining of the recessed 3' OH termini of the proviral DNA at a staggered cut made at the host target site. The fourth step in the process, repair synthesis of the resultant gap, may be accomplished by cellular enzymes.
Nucleotide sequencing of HIV shows the presence of a pol gene in one open reading frame [Ratner, L. et al., Nature, 313, 277(1985)]. Amino acid sequence homology provides evidence that the pol sequence encodes reverse transcriptase, integrase and an HIV protease [Toh, H. et al., EMBO J. 4, 1267 (1985); Power, M. D. et al., Science, 231, 1567 (1986); Pearl, L. H. et al., Nature, 329, 351 (1987)]. All three enzymes have been shown to be essential for the replication of HIV.
It is known that some antiviral compounds which act as inhibitors of HIV replication are effective agents in the treatment of AIDS and similar diseases, e.g., azidothymidine or AZT. Applicants demonstrate that the compounds of this invention are inhibitors of HIV integrase and inhibitors of HIV replication. The applicants additionally demonstrate that inhibition of integrase in vitro and HIV replication in cells is a direct result of inhibiting the strand transfer reaction catalyzed by the recombinant integrase in vitro and integrase as a component of the preintegration complex in HIV infected cells. The particular advantage of the present invention is highly specific inhibition of HIV integrase and HIV replication. The compounds of the present invention inhibit integrases of closely related lentiviruses such as HIV 2 and SIV, but not integrases from more distantly related retroviruses, for example RSV. These compounds do not inhibit binding or catalysis of other nucleic acid binding proteins, including enzymatic reactions such as those catalyzed by HIV reverse transcriptase, HIV Rnase H, Influenza transcriptase, Hepatitis C polymerase, Yeast DNA polymerase, DNase I, Eco RI endonuclease, or mammalian polymerase II.
Zhao et al., (J. Med Chem. vol. 40, pp. 937-941 and 1186-1194 (1997)) describe hydrazide and arylamide FUV integrase inhibitors. Bis-catechols useful for inhibiting HIV integrase are described in LaFemina et al. (Antimicrobial Agents & Chemotherapy, vol. 39, no. 2, pp. 320-324, February 1995).
U.S. Pat. No. 4,871,727 to Burg et al. describes anti-inflammatory and degenerative compounds isolated from the soil microorganism ATCC 20858 of structural formula A below: ##STR2##
wherein:
R is OH or H; PA1 R.sup.1 and R.sup.2 together form .dbd.CH.sub.2 or --CH.sub.2 O--; PA1 R.sup.3 is H, OH, HSO.sub.3 O, HOCOCH.sub.2 CH.sub.2 CO.sub.2 ; PA1 R.sup.4 is OH, HOC.sub.15 H.sub.30 CO.sub.2, AcO or is H except that when R.sup.4 is H, the double bond in the cyclopentane ring is absent; PA1 R.sup.5 is O.dbd., OH, AcO; and PA1 R.sup.6 is OH or --O--C(O)CH.sub.3. PA1 wherein: PA1 "a" is selected from a single bond or a double bond; PA1 R.sup.1 is selected from: PA1 R.sup.2 is selected from: PA1 or R.sup.1 and R.sup.2 are joined to form: ##STR4## PA1 R.sup.3 is selected from: PA1 R.sup.4 is selected from: PA1 R.sup.5 and R.sup.6 are independently selected from: PA1 R.sup.7 is selected from: PA1 or a pharmaceutically acceptable salt thereof. PA1 (a) --OH, PA1 (b) --OC(O)CH.sub.3, PA1 (c) --OC(O)CH.sub.2 C(CH.sub.3)(OH)CH.sub.2 CO.sub.2 H, PA1 (d) --OC(O)CH.sub.2 C(CH.sub.3)(OH)CH.sub.2 CO.sub.2 CH.sub.3, PA1 (e) --OC(O)(CH.sub.2).sub.2 CO.sub.2 H, PA1 (f) --OC(O)(CH.sub.2).sub.2 CO.sub.2 CH.sub.3, PA1 (g) --OC(O)(CH.sub.2).sub.2 CONHOH, PA1 (h) --OCH.sub.2 OCH.sub.3, PA1 (i) --OC(O)C.sub.6 H.sub.5, PA1 (j) --OC(O)CH.sub.2 NH--C(O)OC(CH.sub.3).sub.3, PA1 (k) --OSO.sub.2 CH.sub.3, PA1 (l) --OC(O)CH.sub.2 NH.sub.2, PA1 (m) --OC(O)--(CH.sub.2).sub.15 --OH, and PA1 (n) H. PA1 (a) --OH, PA1 (b) --OC(O)CH.sub.2 C(CH.sub.3)(OH)CH.sub.2 CO.sub.2 H, PA1 (c) --OC(O)CH.sub.2 C(CH.sub.3)(OH)CH.sub.2 CO.sub.2 CH.sub.3, PA1 (d) --OC(O)(CH.sub.2).sub.2 CO.sub.2 H, PA1 (e) --OC(O)(CH.sub.2).sub.2 CONHOH, PA1 (f) --OC(O)CH.sub.2 NH.sub.2, and PA1 (g) --OC(O)--(CH.sub.2).sub.15 --OH. PA1 (a) --OH, PA1 (b) --OC(O)CH.sub.3, PA1 (c) .dbd.O, PA1 (d) --OC(O)(CH.sub.2).sub.2 CO.sub.2 H, PA1 (e) --OC(O)(CH.sub.2).sub.2 CO.sub.2 CH.sub.3, PA1 (f) --OCH.sub.2 OCH.sub.3, PA1 (g) --OC(O)C.sub.6 H.sub.5, PA1 (h) --OC(O)CH.sub.2 NHC(O)OC(CH.sub.3).sub.3, PA1 (i) --OSO.sub.2 OH, and PA1 (j) --OC(O)CH.sub.2 NH.sub.2. PA1 (a) --OH, PA1 (b) .dbd.O, PA1 (c) --OC(O)(CH.sub.2).sub.2 CO.sub.2 H, PA1 (d) --OSO.sub.2 OH, and PA1 (e) --OC(O)CH.sub.2 NH.sub.2. PA1 (a) --H, and PA1 (b) --OH, PA1 or together form: PA1 (c) .dbd.CH.sub.2, or PA1 (d) --CH.sub.2 O--. PA1 (1) an AIDS antiviral agent, PA1 (2) an anti-infective agent, and PA1 (3) an immunomodulator. PA1 (a) fermenting a culture of MF6381 (ATCC 74469), Fusarium sp. or a mutant thereof to produce a fermentation broth, PA1 (b) extracting the fermentation broth with an organic solvent, PA1 (c) isolating the compounds of structural formula I. PA1 (d) chemically modifying the isolated compound of formula I.
Brill et al. (J. Antibiotics, 49(6): 541-546 (1996)), describe particular triterpene sulfates from Fusarium compactum.
PCT publication WO 98/31371 (Application No. PCT/US98/00766) describe the use of androst-5-ene-30-ol-7,17 dione and metabolizable precursors thereof, such as androst-5-ene-3.beta.-acetoxy-7,17-dione, for the treatment of HIV-related weight loss, HIV-related cachexia and HIV-related wasting syndrome.
Applicants have discovered that certain natural product compounds derived from Fusarium sp. MF6381 (ATCC 74469) and derivatives thereof are potent inhibitors of HIV integrase. These compounds are useful for the treatment of AIDS or HIV infections.